Signalling Mechanism in TRPM2-dependent Copper- induced HT22 Cell Death
DOI:
https://doi.org/10.54987/jobimb.v11i2.836Keywords:
Copper toxicity, Oxidative stress, TRPM2 channel, Neuronal death, PARPAbstract
Copper (Cu) is one of the critical elements needed by the human body. However, this metal can cause cytotoxicity when present in excess amounts. In this study, we used HT22 hippocampus cells to examine the function of the Ca2+-permeable transient receptor melastatin 2 (TRPM2) channel in Cu-induced neuronal cell death and the underlying mechanisms. Immunocytochemistry, single-cell imaging, acridine orange/propidium iodide (AO/PI) cell death assay and immunofluorescence microscopy were applied to interpret the mechanisms involved in Cu-induced HT22 cell death. Treatment of 30-300 μM Cu induced an increase in the [Ca2+]i in HT22 cells. Further analysis indicates that Cu exposure induced substantial HT22 cell death. Such response on HT22 cells was significantly inhibited by 2-aminoethoxydiphenyl borate (2-APB) and N-(p-amylcinnamoyl)anthranilic acid (ACA), TRPM2 channel inhibitors. Furthermore, Cu-induced HT22 cell death was suppressed by pharmacologically inhibiting poly(ADPR) polymerase (PARP) using PJ-34 and DPQ. It is known that the activation of TRPM2 channel is via the increase of intracellular reactive oxygen species (ROS). A significant concentration-dependent increase in the generation of ROS was observed in HT22 following the treatment with 30-300 μM Cu. Additionally, Cu-induced HT22 cell death was ablated by inhibiting PKC using CTC, a PKC inhibitor and NADPH oxidase (NOX) using DPI, a NOX generic inhibitor and GKT137831, a NOX1/4-specific inhibitor. Overall, our present study provides evidence suggesting that PKC/NOX/ROS/PARP is an important signalling pathway in Cu-induced activation of the TRPM2 channel and increase in the [Ca2+]i which eventually results in the toxicity of HT22 cells. These results provide new insights into the mechanisms of neurological-associated Cu-induced diseases.
References
Pal A, Jayamani J, Prasad R. An urgent need to reassess the safe levels of copper in the drinking water: Lessons from studies on healthy animals harboring no genetic deficits. Neurotoxicology. 2014;44:58-60.
Chen L, Min J, Wang F. Copper homeostasis and cuproptosis in health and disease. Signal Transduct Target Ther. 2022;7(378).
Jung HS, Kwon PS, Lee JW, Kim JII, Hong CS, Kim JW, et al. Coumarin-derived cu2+-selective fluorescence sensor: Synthesis, mechanisms, and applications in living cells. J Am Chem Soc. 2009;131(5):2008-12.
Frag EYZ, Mohamed ME, Ali AE, Mohamed GG. Potentiometric sensors selective for Cu(II) determination in real water samples and biological fluids based on graphene and MWCNTs modified graphite electrodes. Indian J Chem -Section A. 2020;59(2):162-73.
Lu Y, Yu G, Wei X, Zhan C, Jeon JW, Wang X, et al. Fabric/multi-walled carbon nanotube sensor for portable on-site copper detection in water. Adv Compos Hybrid Mater. 2019;2(4):711-9.
Odetola L, Sills S, Morrison S. A pilot study on the feasibility of testing residential tap water in North Carolina: implications for environmental justice and health. J Expo Sci Environ Epidemiol. 2021;31(6):972-8.
Kirk KA, Andreescu S. Easy-to-Use Sensors for Field Monitoring of Copper Contamination in Water and Pesticide-Sprayed Plants. Anal Chem. 2019;91(21):13892-9.
Zheng W, Monnot AD. Regulation of brain iron and copper homeostasis by brain barrier systems: Implication in neurodegenerative diseases. Pharmacol Ther. 2012;133(2):177-88.
Ackerman CM, Chang CJ. Copper signaling in the brain and beyond. J Biol Chem. 2018;293(13):4628-35.
An Y, Li S, Huang X, Chen X, Shan H, Zhang M. The Role of Copper Homeostasis in Brain Disease. Int J Mol Sci. 2022;23(22):1-23.
Liao J, Yang F, Chen H, Yu W, Han Q, Li Y, et al. Effects of copper on oxidative stress and autophagy in hypothalamus of broilers. Ecotoxicol Environ Saf. 2019;185(July).
Li M, Cui W, Peng X, Bai C, Cui H. Effect of dietary high copper on the oxidization in brain tissue of chickens. Chinese J Vet Sci. 2009;29(10):1334-7 ref.14.
Li M, Cui W, Peng X, Bai C, Cui H. Effect of dietary high copper on the antioxidase activities of brain tissue in chickens. Acta Vet Zootech Sin. 2010;41(2)(220-3).
Morris MC, Evans DA, Tangney CC, Bienias JL, Schneider JA, Wilson RS, et al. Dietary copper and high saturated and trans fat intakes associated with cognitive decline. Arch Neurol. 2006;63(8):1085-8.
Ventriglia M, Bucossi S, Panetta V, Squitti R. Erratum: Copper in alzheimer's disease: A meta-analysis of serum, plasma, and cerebrospinal fluid studies (Journal of Alzheimer's Disease (2012) 30 (9981-984)). J Alzheimer's Dis. 2012;30(4):981-4.
Przyby?kowski A, Gromadzka G, Wawer A, Bulska E, Jab?onka-Salach K, Grygorowicz T, et al. Neurochemical and behavioral characteristics of toxic milk mice: An animal model of wilson's disease. Neurochem Res. 2013;38(10):2037-45.
Squitti R, Ghidoni R, Siotto M, Ventriglia M, Benussi L, Paterlini A, et al. Value of serum nonceruloplasmin copper for prediction of mild cognitive impairment conversion to Alzheimer disease. Ann Neurol. 2014;75(4):574-80.
Alak G, Ucar A, Yeltekin AÇ, Çomakl? S, Parlak V, Ta? IH, et al. Neuroprotective effects of dietary borax in the brain tissue of rainbow trout (Oncorhynchus mykiss) exposed to copper-induced toxicity. Fish Physiol Biochem. 2018;44(5):1409-20.
Su R, Wang R, Guo S, Cao H, Pan J, Li C, et al. In vitro effect of copper chloride exposure on reactive oxygen species generation and respiratory chain complex activities of mitochondria isolated from broiler liver. Biol Trace Elem Res. 2011;144(1-3):668-77.
Wang Y, Zhao H, Shao Y, Liu J, Li J, Luo L, et al. Copper (II) and/or arsenite-induced oxidative stress cascades apoptosis and autophagy in the skeletal muscles of chicken. Chemosphere. 2018;206:597-605.
Auten RL, Davis JM. Oxygen toxicity and reactive oxygen species: The devil is in the details. Pediatr Res. 2009;66(2):121-7.
Salim S. Oxidative stress and the central nervous system. J Pharmacol Exp Ther. 2017;360(1):201-5.
Nunes KZ, Fioresi M, Marques VB, Vassallo DV. Acute copper overload induces vascular dysfunction in aortic rings due to endothelial oxidative stress and increased nitric oxide production. J Toxicol Environ Heal - Part A Curr Issues. 2018;81(8):218-28.
Yang F, Liao J, Pei R, Yu W, Han Q, Li Y, et al. Autophagy attenuates copper-induced mitochondrial dysfunction by regulating oxidative stress in chicken hepatocytes. Chemosphere. 2018;204:36-43.
Shimada K, Reznik E, Stokes ME, Krishnamoorthy L, Bos PH, Song Y, et al. Copper-Binding Small Molecule Induces Oxidative Stress and Cell-Cycle Arrest in Glioblastoma-Patient-Derived Cells. Cell Chem Biol. 2018;25(5):585-594.e7.
Rakshit A, Khatua K, Shanbhag V, Comba P, Datta A. Cu2+ selective chelators relieve copper-induced oxidative stress: In vivo. Chem Sci. 2018;9(41):7916-30.
Alawieyah Syed Mortadza S, Sim JA, Neubrand VE, Jiang LH. A critical role of TRPM2 channel in A?42-induced microglial activation and generation of tumor necrosis factor-?. Glia. 2018;66(3):562-75.
Yeh FL, Hansen D V, Sheng M. Focus Issue?: Feature Review TREM2, Microglia, and Neurodegenerative Diseases. Trends Mol Med. 2017;23(6):512-33.
Ferreira AFF, Singulani MP, Ulrich H, Feng Z-P, Sun H-S, Britto LR. Inhibition of TRPM2 by AG490 Is Neuroprotective in a Parkinson's Disease Animal Model. Mol Biol. 2022;59(3):1543-1559.
Ali ES, Chakrabarty B, Ramproshad S, Mondal B, Kundu N, Sarkar C, et al. TRPM2-mediated Ca2+ signaling as a potential therapeutic target in cancer treatment: an updated review of its role in survival and proliferation of cancer cells. Cell Commun Signal. 2023;21(1):1-13.
Mortadza SAS, Wang L, Li D, Jiang LH. TRPM2 channel-mediated ROS-sensitive Ca2+ signaling mechanisms in immune cells. Front Immunol. 2015;6(JUL):1-7.
Olah ME. the Activation of Novel Calcium-Dependent Pathways Downstream of N-Methyl-D- Aspartate Receptors. 2009.
Bai JZ, Lipski J. Differential expression of TRPM2 and TRPV4 channels and their potential role in oxidative stress-induced cell death in organotypic hippocampal culture. Neurotoxicology. 2010;31(2):204-14.
Li X, Yang W, Jiang LH. Alteration in intracellular Zn2+ homeostasis as a result of TRPM2 channel activation contributes to ROS-induced hippocampal neuronal death. Front Mol Neurosci. 2017;10(December):1-11.
Li X, Jiang LH. Multiple molecular mechanisms form a positive feedback loop driving amyloid ?42 peptide-induced neurotoxicity via activation of the TRPM2 channel in hippocampal neurons article. Cell Death Dis. 2018;9(2):1-16.
Xu X, Chua CC, Zhang M, Geng D, Liu CF, Hamdy RC, et al. The role of PARP activation in glutamate-induced necroptosis in HT-22 cells. Brain Res. 2010;1343:206-12.
Xu H, Luo P, Zhao Y, Zhao M, Yang Y, Chen T, et al. Iduna protects HT22 cells from hydrogen peroxide-induced oxidative stress through interfering poly(ADP-ribose) polymerase-1-induced cell death (parthanatos). Cell Signal. 2013;25(4):1018-26.
Wang X, Zhang W, Ge P, Yu M, Meng H. Parthanatos participates in glutamate-mediated HT22 cell injury and hippocampal neuronal death in kainic acid-induced status epilepticus rats. CNS Neurosci Ther. 2022;28(12):2032-43.
Ying Y, Gong L, Tao X, Ding J, Chen N, Yao Y, et al. Genetic Knockout of TRPM2 Increases Neuronal Excitability of Hippocampal Neurons by Inhibiting Kv7 Channel in Epilepsy. Mol Neurobiol. 2022;(59):6918-6933.
Danese A, Leo S, Rimessi A, Wieckowski MR, Fiorica F, Giorgi C, et al. Cell death as a result of calcium signaling modulation: A cancer-centric prospective. Biochim Biophys Acta - Mol Cell Res. 2021;1868(8).
Malko P, Jiang LH. TRPM2 channel-mediated cell death: An important mechanism linking oxidative stress-inducing pathological factors to associated pathological conditions. Redox Biol. 2020;37.
Wang X, Xiao Y, Huang M, Shen B, Xue H, Wu K. Effect of TRPM2-Mediated Calcium Signaling on Cell Proliferation and Apoptosis in Esophageal Squamous Cell Carcinoma. Technol Cancer Res Treat. 2021;20:1-8.
Sha'fi MSA, Rathakrishnan S, Hazanol IN, Izzazuddin, Dali MHI, Khayat ME, et al. Ethanol Induces Microglial Cell Death via the. Antioxidants. 2020;9(1253):1-11.
Ganguly U, Kaur U, Chakrabarti SS, Sharma P, Agrawal BK, Saso L, et al. Oxidative Stress, Neuroinflammation, and NADPH Oxidase: Implications in the Pathogenesis and Treatment of Alzheimer's Disease. Oxid Med Cell Longev. 2021;2021.
Hernandes MS, Xu Q, Griendling KK. Role of NADPH Oxidases in Blood-Brain Barrier Disruption and Ischemic Stroke. Antioxidants. 2022;11(10).
Terzi A, Suter DM. The role of NADPH oxidases in neuronal development. Free Radic Biol Med. 2020;154(May):33-47.
Lee SH, Lee MW, Ko DG, Choi BY, Suh SW. The role of nadph oxidase in neuronal death and neurogenesis after acute neurological disorders. Antioxidants. 2021;10(5):1-14.
Mortadza SS, Sim JA, Stacey M, Jiang LH. Signalling mechanisms mediating Zn 2+ -induced TRPM2 channel activation and cell death in microglial cells. Sci Rep. 2017;7(February):1-15.
Rastogi R, Geng X, Li F, Ding Y. NOX activation by subunit interaction and underlying mechanisms in disease. Front Cell Neurosci. 2017;10(January):1-14.
Jin Q, Jiang Y, Fu L, Zheng Y, Ding Y, Liu Q. Wenxin Granule Ameliorates Hypoxia/Reoxygenation-Induced Oxidative Stress in Mitochondria via the PKC- ? /NOX2/ROS Pathway in H9c2 Cells. Oxid Med Cell Longev. 2020;2020.
Kim SG, Sung JY, Kang YJ, Choi HC. Fisetin alleviates cellular senescence through PTEN mediated inhibition of PKC?-NOX1 pathway in vascular smooth muscle cells. Arch Gerontol Geriatr. 2023;108(January).
Lin MC, Liu CC, Lin YC, Liao CS. Resveratrol protects against cerebral ischemic injury via restraining lipid peroxidation, transition elements, and toxic metal levels, but enhancing anti-oxidant activity. Antioxidants. 2021;10(10).
Timoshnikov VA, Selyutina OY, Polyakov NE, Didichenko V, Kontoghiorghes GJ. Mechanistic Insights of Chelator Complexes with Essential Transition Metals: Antioxidant/Pro-Oxidant Activity and Applications in Medicine. Int J Mol Sci. 2022;23(3).
Ranjan A, Kalita J, Kumar V, Misra UK. MRI and oxidative stress markers in neurological worsening of Wilson disease following penicillamine. Neurotoxicology. 2015;49:45-9.
Hermann W. Cerebral Imaging in Wilson Disease. In: Cinical and Translational Perspectives on Wilson Disease. Academic Press; 2019. p. 271-8.
Radovanovi? V, Vlaini? J, Hanži? N, Uki? P, Oršoli? N, Baranovi? G, et al. Neurotoxic effect of ethanolic extract of propolis in the presence of copper ions is mediated through enhanced production of ROS and stimulation of caspase-3/7 activity. Toxins (Basel). 2019;11(5).
Sun Y, Sukumaran P, Selvaraj S, Cilz NI, Schaar A, Lei S, et al. TRPM2 Promotes NeurotoxinMPP /MPTP-Induced Cell Death. Mol Biol. 2018;(55):409-20.
Li X, Jiang LH. A critical role of the transient receptor potential melastatin 2 channel in a positive feedback mechanism for reactive oxygen species-induced delayed cell death. J Cell Physiol. 2019;234(4):3647-60.
An X, Fu Z, Mai C, Wang W, Wei L, Li D, et al. Increasing the TRPM2 channel expression in human neuroblastoma SH-SY5Y Cells augments the susceptibility to ROS-induced cell death. Cells. 2019;8(1).
Güzel M, Naz?ro?lu M, Akp?nar O, Ç?nar R. Interferon Gamma-Mediated Oxidative Stress Induces Apoptosis, Neuroinflammation, Zinc Ion Influx, and TRPM2 Channel Activation in Neuronal Cell Line: Modulator Role of Curcumin. Inflammation. 2021;44(5):1878-94.
Jung S, Chung Y, Lee Y, Lee Y, Cho JW, Shin EJ, et al. Buffering of cytosolic calcium plays a neuroprotective role by preserving the autophagy-lysosome pathway during MPP+-induced neuronal death. Cell Death Discov. 2019;5(1).
Calvo-rodriguez M, Hou SS, Snyder AC, Kharitonova EK, Russ AN, Das S, et al. Increased mitochondrial calcium levels associated with neuronal death in a mouse model of Alzheimer's disease. Nat Commun. 2020;11:2146
Guan PP, Cao LL, Yang Y, Wang P. Calcium Ions Aggravate Alzheimer's Disease Through the Aberrant Activation of Neuronal Networks, Leading to Synaptic and Cognitive Deficits. Front Mol Neurosci. 2021;14(December):1-32.
Reddy PVB, Rama Rao K V., Norenberg MD. The mitochondrial permeability transition, and oxidative and nitrosative stress in the mechanism of copper toxicity in cultured neurons and astrocytes. Lab Investig. 2008;88(8):816-30.
Jazvinš?ak Jembrek M, Vlaini? J, Radovanovi? V, Erhardt J, Oršoli? N. Effects of copper overload in P19 neurons: Impairment of glutathione redox homeostasis and crosstalk between caspase and calpain protease systems in ROS-induced apoptosis. BioMetals. 2014;27(6):1303-22.
Wandt VK, Winkelbeiner N, Bornhorst J, Witt B, Raschke S, Simon L, et al. A matter of concern - Trace element dyshomeostasis and genomic stability in neurons. Redox Biol. 2021;41(December 2020).
Gonzalez-Alcocer A, Gopar-Cuevas Y, Soto-Dominguez A, Castillo-Velazquez U, de Jesus Loera-Arias M, Saucedo-Cardenas O, et al. Combined chronic copper exposure and aging lead to neurotoxicity in vivo. Neurotoxicology. 2023;95(February):181-92.
Togashi K , Inada H, and Tominaga M. Inhibiton of the transient receptor potential cation channel TRPM2 by 2-aminoethoxydiphenyl borate (2-APB). Br. J. Pharmacology. 2008; 153 (6): 1324-1330
Kühn, F.J.P., Mathis, W., Cornelia, K. et al. Modulation of activation and inactivation by Ca2+ and 2-APB in the pore of an archetypal TRPM channel from Nematostella vectensis . Sci Rep. 2017; 7: 7245.
Özge K., Yurdun K., Bilge G., Ebru D., Yenilmez et al. Investigation the effects of 2-aminoethoxydiphenyl borate (2-APB) on aminoglycoside nephrotoxicity. Ultrastructural Pathology. 2023; 3: 121-128
Kraft R, Grimm C, Frenzel H, Harteneck C. Inhibition of TRPM2 cation channels by N-(p-amylcinnamoyl)anthranilic acid. Br J Pharmacol. 2006 Jun;148(3):264-73.
Luo Y, Yu X, Ma C, Luo J and Yang W. Identification of a Novel EF-Loop in the N-terminus of TRPM2 Channel Involved in Calcium Sensitivity. Frontier in Pharmacology. 2018; 9:581.
Jiang LH, Yang W, Zou J, Beech DJ. TRPM2 channel properties, functions and therapeutic potentials. Expert Opin Ther Targets. 2010;14(9):973-88.
Dai C, Liu Q, Li D, Sharma G, Xiong J, Xiao X. Molecular insights of copper sulfate exposure-induced nephrotoxicity: Involvement of oxidative and endoplasmic reticulum stress pathways. Biomolecules. 2020;10(7):1-15.
Zhou Q, Zhang Y, Lu L, Zhang H, Zhao C, Pu Y, et al. Copper induces microglia-mediated neuroinflammation through ROS/NF-?B pathway and mitophagy disorder. Food Chem Toxicol. 2022;168(August).
Noh KM, Koh JY. Induction and activation by zinc of NADPH oxidase in cultured cortical neurons and astrocytes. J Neurosci. 2000;20(23):1-5.
Koh JY. Zinc and disease of the brain. Mol Neurobiol. 2001;24(1-3):99-106.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Journal of Biochemistry, Microbiology and Biotechnology
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
